Smart Protein Layers (SPL) is a standard-based technology for stain-free, quantitative and standardized analysis of proteins separated by 1D gel electrophoresis. SPL is based on rapid fluorescent labeling of protein samples using Smart Labels and a bi-fluorescent standard, the Smartalyzer (SMA). The relation between fluorescent labeled protein sample and the SMA allows highly sensitive, stain-free protein visualization as well as for precise protein normalization and quantification.

In this paper we demonstrate that the SPL technology allows for absolute protein quantification even if varying amounts of protein have been applied to the gel - either by variations in the protein content or by loading different amounts of sample.

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The patented SPL technology is designed for stain-free, qualitative and quantitative protein analysis in protein gels ... learn more here

... as well as to co-detect sample protein on Western Blots while the target is detected (reliable Western Blot normalization) ... learn more here.

Real-time Normalization of Western Blots: Smart Protein Layers

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Over decades, Western blotting is a method to detect a target protein in a complex sample. The technique includes protein separation, transfer to a membrane and several washing and antibody incubation steps.
Reliable comparison of detected target signal intensities between different samples is still a problem. It requires an appropriate way of normalization to assure that the reported fold changes of the target protein are not an artifact of a reference signal and/ or experimental errors.

Smart Protein Layers (SPL) is a add-on kit for the detection of sample proteins present on the blot at the same time the target is immuodetected. It is based on two components: i) fluorescent SPL Labels, bound to the sample protein prior separation, for the visualization of all proteins present on the blot at this time ii) bi-fluorescent SPL Standards to monitor labeling efficiency and to provide additional information about the sample and to enable automated data evaluation.

With SPL the determined expression of a specific target protein can be normalized to its corresponding sample, providing a reliable and precise tool for the comparison of fold changes between different samples.